2009
DOI: 10.1126/science.1176325
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C3PO, an Endoribonuclease That Promotes RNAi by Facilitating RISC Activation

Abstract: The catalytic engine of RNAi is the RNA-induced silencing complex (RISC), wherein the endoribonuclease Argonaute and single-stranded siRNA direct target mRNA cleavage. Here we have reconstituted long dsRNA- and duplex siRNA-initiated RISC activities using recombinant Drosophila Dicer-2, R2D2 and Ago2 proteins. We employ this core reconstitution system to purify an RNAi regulator-component 3 promoter of RISC (C3PO), a complex of Translin and Trax. C3PO is a Mg2+-dependent endoribonuclease that promotes RISC act… Show more

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Cited by 229 publications
(269 citation statements)
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“…Our mass spectrometry analyses support this view, because we did not find proteins that participate in small RNA biogenesis or RISC assembly. We detected no peptides from Dicer-1, Dicer-2, R2D2, Loquacious, or C3PO (Jiang et al 2005;Liu et al 2006Liu et al , 2007Liu et al , 2009) copurifying with Drosophila Ago1-or Ago2-RISC, nor did we detect Dicer, TRBP, or PACT (Hutvágner et al 2001;Chendrimada et al 2005;Haase et al 2005;Lee et al 2006) copurifying with mouse AGO2-RISC (Supplemental Fig. S3; Supplemental Tables S1-S3).…”
Section: Discussionmentioning
confidence: 91%
“…Our mass spectrometry analyses support this view, because we did not find proteins that participate in small RNA biogenesis or RISC assembly. We detected no peptides from Dicer-1, Dicer-2, R2D2, Loquacious, or C3PO (Jiang et al 2005;Liu et al 2006Liu et al , 2007Liu et al , 2009) copurifying with Drosophila Ago1-or Ago2-RISC, nor did we detect Dicer, TRBP, or PACT (Hutvágner et al 2001;Chendrimada et al 2005;Haase et al 2005;Lee et al 2006) copurifying with mouse AGO2-RISC (Supplemental Fig. S3; Supplemental Tables S1-S3).…”
Section: Discussionmentioning
confidence: 91%
“…After nicking, the endonuclease component 3 promoter of RISC complex (C3PO), composed of Trax and Translin proteins in humans, is able to cleave and remove the passenger strand [87,90,91] . This results in activation of pre-RISC into the RISC complex.…”
Section: Ptgsmentioning
confidence: 99%
“…Following AGO2 loading, the duplex strand with the less stable 5′ terminus (the guide strand) is selectively retained, whereas the complementary (passenger) strand is cleaved by the slicer activity of AGO2, a process known as passenger-strand nicking (19)(20)(21)(22). The siRNA duplex is subsequently unwound, and the sliced passenger strand is removed by the C3PO endoribonuclease complex (23), resulting in the formation of the mature RISC that contains the siRNA guide strand. Finally, target mRNAs are recruited to mature RISC by perfectly complementary base pairing with the siRNA guide strand and cleaved (sliced) by AGO2 (24,25).…”
mentioning
confidence: 99%